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[Preprint]. 2024 May 15:2023.11.03.565545. Originally published 2023 Nov 3. [Version 2] doi: 10.1101/2023.11.03.565545

Translation initiation factor eIF1.2 promotes Toxoplasma stage conversion by regulating levels of key differentiation factors

Fengrong Wang, Michael J Holmes, Hea Jin Hong, Pariyamon Thaprawat, Geetha Kannan, My-Hang Huynh, Tracey L Schultz, M Haley Licon, Sebastian Lourido, Wenzhao Dong, Jailson Brito Querido, William J Sullivan, Seán E O’Leary, Vern B Carruthers
PMCID: PMC10635126  PMID: 37961607

Abstracts

The parasite Toxoplasma gondii persists in its hosts by converting from replicating tachyzoites to latent bradyzoites housed in tissue cysts. The molecular mechanisms that mediate T. gondii differentiation remain poorly understood. Through a mutagenesis screen, we identified translation initiation factor eIF1.2 as a critical factor for T. gondii differentiation. A F97L mutation in eIF1.2 or the genetic ablation of eIF1.2eif1.2 ) markedly impeded bradyzoite cyst formation in vitro and in vivo . We demonstrated, at single-molecule level, that the eIF1.2 F97L mutation impacts the scanning process of the ribosome preinitiation complex on a model mRNA. RNA sequencing and ribosome profiling experiments unveiled that Δ eif1.2 parasites are defective in upregulating bradyzoite induction factors BFD1 and BFD2 during stress-induced differentiation. Forced expression of BFD1 or BFD2 significantly restored differentiation in Δ eif1.2 parasites. Together, our findings suggest that eIF1.2 functions by regulating the translation of key differentiation factors necessary to establish chronic toxoplasmosis.

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